LOCUS       sequence-488218-        7338 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_201, derived from human fusion protein TSG101_YWHAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7338)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7338)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_201
FEATURES             Location/Qualifiers
     source          1..7338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..200
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    199..254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     300..319
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(871..1581,1582..1584,1585..1587))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(913..934)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1262..1281
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1521..1542
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1584..1593
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1669..1872)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1698..1718)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1873..2176
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2433..2477)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2662..2895
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    3255..3372
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3864..3989
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4036..4293
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4295..4674
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4705..4809
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4810..4878,4879..5670)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5028..5047)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5841..6429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6330..6349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6583..6600
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6668..6802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6692..6711)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6746..6765
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6808..7003)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6822..6957
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7062..7083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7098..7103,7104..7121,7122..7128)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7136..7152
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7141..7163
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7160..7176
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7160..7176
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
       61 ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg
      121 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
      181 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
      241 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
      301 tttggcagag ggaaaaagat ctgctagctc gagcagctga agctttattg aggcttaagc
      361 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
      421 ccagtacagg caaaaagcag atcttgtctt ctttgggagt gaattagccc ttccagtccc
      481 cccttttctt ttaaaaagtg gctaagatct acagctgcct tgtaagtcat tggtcttaaa
      541 ggtaccgtaa tcttatctag aaccggttca ctcgatcttc tcgcggtact cttttcccat
      601 ttgctgcttt ttttcattac gttctgtttc gactgttgta accggtggtt gggaggggta
      661 ttgtgaggat gtggttgcag ggtatggccc accgggaggg taagggcaac caggatagcc
      721 agaagggtta ggagggtaac ctgacgggta tgggctaatg ccgcccggca taccgggcat
      781 gtatgacgta tttggcggcc cggtagcctg atacggcggg taggaagccc gggaagcggc
      841 cgcacttccc ccggagcctc cgcttccgcc cttgtacagc tcgtccatgc cgagagtgat
      901 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
      961 gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
     1021 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg
     1081 gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt
     1141 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     1201 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     1261 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
     1321 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg
     1381 cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     1441 gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
     1501 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     1561 gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt ggatcccggg cccgcggtac
     1621 cgtcgactgc agaattcggt tcactaaacg agcccggttc actaaacgag ctctgcttat
     1681 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     1741 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     1801 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     1861 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     1921 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     1981 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     2041 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     2101 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     2161 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     2221 ttactattaa taactagcac gcgttcgaag cttgagctcg gtcccctcgg ggttgggagg
     2281 tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca
     2341 gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac
     2401 cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat
     2461 aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag
     2521 cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt
     2581 tccagagatt tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt
     2641 ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg
     2701 attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc
     2761 tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc
     2821 tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca
     2881 aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc
     2941 ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata taattcactt
     3001 ctccaattgt ccctcatatc tcctccttat ggtgttttac taaacttttc catgttctaa
     3061 tcctcatcct gtctacttgc cacacaatca tcacctgcca tctgttttcc ataatcccta
     3121 atgatctttg cttttcttct tggcactact tttatgtcac tattatcttg tattactact
     3181 gccccttcac ctttccagag gagctttgct ggtcctttcc aaagtggatt tctgctgtcc
     3241 ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc tttagtttgt
     3301 atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc aatcccccct
     3361 tttcttttaa aattgtggat gaatactgcc atttgtactg ctgtcttaag atgttcagcc
     3421 tgatctctta cctgtcctat aattttcttt aattctttat tcatagattc tactactcct
     3481 tgactttggg gattgtaggg aatttccagg tctgaagatc ctgctgcttg ctgtgctttt
     3541 ttcttacttt tgttttgctc ttcctctatc ttgtctaaag cttccttggt gtcttttatc
     3601 tctatccttt gatgcacaca atagagggtt gctactgtat tatataatga tctaagttct
     3661 tctgatcctg tctgaaggga tggttgtagc tgtcccagta tttgtctaca gccttctgat
     3721 gtttctaaca ggccagtatt aactgcgaat cgttctagct ccctgcttgc ccatactata
     3781 tgttttaatt tatatttttt ctttccccct ggccttaacc gaattttttc ccatcgcgat
     3841 ctaattctcc cccgcttaat actgacgctc tcgcacccat ctctctcctt ctagcctccg
     3901 ctagtcaaaa tttttggcgt actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct
     3961 tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt tccctttcgc tttcaagtcc
     4021 ctgttcgggc gccactgcta gagattttcc acactgacta aaagggtctg agggatctct
     4081 agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt
     4141 attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt
     4201 ctaaccagag agacccagta caggcaaaaa gcagctgctt atatgcagga tctgagggct
     4261 cgccactccc cagtcccgcc caggccacgc ctcccatggt aatagcgatg actaatacgt
     4321 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     4381 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     4441 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     4501 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     4561 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     4621 gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgtcgaccca
     4681 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4741 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4801 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4861 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4921 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4981 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5041 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5101 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5161 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5221 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5281 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5341 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5401 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5461 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5521 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5581 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5641 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5701 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5761 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5821 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5881 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5941 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6001 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6061 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6121 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6181 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6241 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6301 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6361 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6421 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6481 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6541 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6601 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6661 tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     6721 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     6781 tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     6841 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     6901 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     6961 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcggatccg ctggcacgac
     7021 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7081 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7141 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgggctg
     7201 caggtcgagg gatctccata agagaagagg gacagctatg actgggagta gtcaggagag
     7261 gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa
     7321 taacacaaaa caaaatat
//
